Crystalline form of r)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin- 5-yl)-3-fluorophenyl)-5-hydroxymethyl oxazolidin-2-one dihydrogen phosphate

ABSTRACT

A crystalline form of crystalline (R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyl oxazolidin-2-one dihydrogen phosphate, methods of making the crystalline form and pharmaceutical compositions comprising the crystalline form are useful antibiotics. Further, the derivatives of the present invention may exert potent antibacterial activity versus various human and animal pathogens, including Gram-positive bacteria such as Staphylococi, Enterococci and Streptococi, anaerobic microorganisms such as  Bacteroides  and Clostridia, and acid-resistant microorganisms such as  Mycobacterium tuberculosis  and  Mycobacterium avium . Accordingly, the compositions comprising the crystalline form may be used in antibiotics.

This application is a Continuation of U.S. patent application Ser. No.12/699,864, filed Feb. 3, 2010, which claims priority to U.S.Provisional Patent Application No. 61/149,402, filed Feb. 3, 2009, thefull disclosure of each of these documents is incorporated herein byreference in its entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present disclosure relates to a crystalline form of(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate, and methods of making and usingthe crystalline form. The crystalline form may be used as apharmaceutically active compound in compositions that are useful inimpeding the growth of bacteria or treating patients suffering frombacterial infections.

2. Description of the Related Art

US Patent Publication No. 20070155798, which is hereby incorporated byreference in its entirety, recently disclosed a series of potentlyanti-bacterial oxazolidinones including

wherein R═H, PO(OH)₂, and PO(ONa)₂.

Although this patent application discloses methods of making compoundssuch as the free acid (wherein R═PO(OH)₂) and the disodium salt (whereinR═PO(ONa)₂), there is no indication that any of the compounds werestably crystallized or purified. In addition, these processes includethe use of reagents which are highly corrosive, such as trichloroaceticacid, or explosive, such as ethyl ether, and therefore are not suitablefor commercial use. As discussed below in more detail, attempts tocrystallize the disodium salt by the instant inventors resulted in ahighly hygroscopic, unstable crystalline salt form which turnedamorphous upon drying.

There is a need in the art for a stable, non-hygroscopic crystallineform of the free acid (wherein R═PO(OH)₂) or a salt thereof that can beaccurately poured and weighed for use in pharmaceutical formulations.Also, it would be advantageous if the crystalline form did not form alarge number of polymorphs, as the number of polymorphs hinders theability to reproducibly provide the identical polymorph duringmanufacturing. Making a particular crystalline form having theseproperties is an empirical process, and one skilled in the art would beunable to predict among the free acid form of the pharmaceuticalcompound or one of the corresponding salts, which would crystallize, ifat all, under which crystallization conditions. In addition, one skilledin the art would be unable to predict which crystalline form would havethe beneficial properties of stability, pourability, non-hygroscopicityand reproducibility.

In addition, improved methods of making the free acid are disclosed inU.S. patent application Ser. No. 12/577,089, which is assigned to TriusTherapeutics, Inc., and which is incorporated herein by reference.Difficulties in filtering crystalline material and processing thecrystalline material into dosage forms, such as tablets, have arisenbecause the free acid forms fine particles which delay processing time.Therefore, there is also a need in the art for a crystalline form of thecompound and related methods that overcome these processingdifficulties.

In addition, it would be advantageous to have a purified compound thatis suitable for pharmaceutical compositions.

SUMMARY OF THE INVENTION

Surprisingly, a crystalline(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂), was more stable andnon-hygroscopic than the salt forms that were tested. In addition,unlike typical crystallizations, where the crystallization conditions,such as the solvent and temperature conditions, determine the particularcrystalline form, the same crystalline form of 1 (R═PO(OH)₂) wasproduced using many solvent and crystallization conditions. Therefore,this crystalline form was very stable, was made reproducibly, and idealfor commercial production because it reduced the chances that otherpolymorphs would form contaminating impurities during production.However, in all preliminary testing, the free acid crystallized as fineparticles, making filtering and processing difficult.

To overcome difficulties in filtering and processing crystalline(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂), processes describedherein result in significantly reduced filtering time, avoid more toxicsolvents, and significantly increased ease of preparing dosage formssuch as tablets. It has been found that implementing various processescan control the particle size distribution of the resulting material,which is useful for making the crystalline form, and for commercialproduction and pharmaceutical use. Surprisingly, the process forincreasing the particle size reduces the amount of the dimer impurity,in comparison to the process for making the free acid disclosed in U.S.patent application Ser. No. 12/577,089. Thus, various methods of makingand using the crystalline form are also provided.

In addition, by using methods of making the free acid disclosed in U.S.patent application Ser. No. 12/577,089, which is assigned to the sameassignee as in the present application, and by using the crystallizationmethods described herein, a crystalline free acid having at least 96%purity by weight may be formed that comprises a compound having thefollowing formula:

(hereinafter “the chloro impurity”), i.e.,(R)-5-(chloromethyl)-3-(3-fluoro-4-(6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl)phenyl)oxazolidin-2-onein an amount less than 1%.

Similarly, by using methods of making the free acid disclosed in U.S.patent application Ser. No. 12/577,089, which is assigned to the sameassignee as in the present application, and by using the crystallizationmethods described herein, a crystalline free acid having at least 96%purity by weight may be formed that comprises a compound having thefollowing formula:

(hereinafter “TR-700”), i.e., 5R)-3-{3-Fluoro-4-[6-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)-pyridin-3-yl]-phenyl}-5-hydroxymethyl-1,3-oxazolidin-2-one,in an amount less than 1%.

The crystalline free acid may have one or more of the attributesdescribed herein.

In some aspects, a purified crystalline(R)-3-(4-(2-(2-methyltetrazol-5-yl)-pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate, i.e., the free acid, has a purityof at least about 96% by weight. In some embodiments, the crystallinefree acid has a median volume diameter of at least about 1.0 μm.

In some embodiments, pharmaceutical composition comprises the free acidor a salt thereof and at least one pharmaceutically acceptable carrier,excipient or diluent.

In some embodiments, a method of treating a bacterial infectioncomprises administering an effective amount of the crystalline freeacid, or a salt thereof to a subject in need thereof. Methods may alsoinclude comprise treating a bacterial infection comprising administeringthe free acid, pharmaceutical composition thereof or a salt to a subjectin need thereof.

In some aspects, processes for making the free acid comprise dryingcrystallized(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate, or a pharmaceutical compositioncomprising the salt thereof.

These and other embodiments are described in greater detail below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 the FT-Raman spectrum of crystalline 1 (R═PO(OH)₂).

FIG. 2 shows the X-ray powder pattern of crystalline 1 (R═PO(OH)₂).

FIG. 3 shows the differential scanning calorimetry (DSC) thermogram ofcrystalline 1 (R═PO(OH)₂).

FIGS. 4-1 to 4-3 show the ¹-H NMR spectrum of 1 (R═PO(OH)₂).

FIG. 5 depicts the TG-FTIR diagram of crystalline 1 (R═PO(OH)₂).

FIG. 6 is a diagram showing the dynamic vapor sorption (DVS) behavior ofcrystalline 1 (R═PO(OH)₂).

FIG. 7 is a manufacturing process schematic for 1 (R═PO(OH)₂) (TR-701FA) in a tablet dosage form.

FIG. 8 is a manufacturing process schematic for 1 (R═PO(OH)₂) (TR-701FA) Compounding Solution for Lyophilization.

FIG. 9 is a manufacturing process schematic for 1 (R═PO(OH)₂) (TR-701FA) for Injection, 200 mg/vial: sterile filtering, filling, andlyophilization.

FIG. 10 is a representative particle size distribution of crystallinefree acid without regard to controlling particle size distribution asalso described herein.

FIG. 11 is a representative particle size distribution of crystallinefree acid made using laboratory processes to control particle sizedescribed herein.

FIG. 12 is a representative particle size distribution of crystallinefree acid made using scaled up manufacturing processes to controlparticle size described herein.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂), which is sometimesreferred to herein as the “free acid” or “TR-701 FA,” and several saltsthereof were prepared under various crystallization conditions todetermine which of the materials would form the most stable and leasthygroscopic crystalline compound. The empirical process of makingcrystalline forms of the free acid and salts thereof resulted in theselection of a crystalline free acid that, in addition to superiorstability and non-hygroscopicity, was reproducibly made under variouscrystallization conditions, which was subsequently purified and dried.

Specifically, most of the salts that were evaluated were difficult toprepare in a crystalline form or were otherwise unstable, such as in apurified or dried form. For example, with respect to the mono-sodiumsalt, the formation of a stable hydrate was not detected. Also, thematerial contained over 10% by weight of water and therefore thematerial was very hygroscopic, and thus not suitable for the desireduse.

A disodium salt crystalline hydrate was formed, but was unstable andcontained 19.6% by weight of water. The disodium salt, however, was verysoluble. Drying the hydrate resulted in amorphous samples. The watercontent of an amorphous sample was about 6.2% by weight.

A crystalline solid material was not isolated for a di-potassium.

A hemi-calcium salt was prepared as a crystal, however, it wasunsuitably hygroscopic.

A hemi-magnesium salt crystalline material was formed and appeared tocontain various hydrates of a salt, and therefore, the presence ofvarious polymorphs would render it less desirable for use in aformulation. In one experiment, a magnesium salt had a melting point of152.8° C., which in this case indicated that this material was lessstable in comparison to the free acid.

The free acid formed crystals, which were non-hygroscopic upon filteringand drying, which showed an aqueous solubility of 0.1 mg/ml (pH=3.2 ofthe saturated solution). The crystalline material's melting point wasapproximately 255-258° C., and therefore was very stable at a relativelyhigh temperature.

Generally, the crystallization conditions are usually critical forforming a particular polymorph; however, surprisingly, the same freeacid polymorph was formed under all of the various conditions in whichthe crystalline free acid was formed.

In some embodiments, the crystalline material is non-hygroscopic, so itdoes not readily take up and retain water from the atmosphere. In someembodiments, “non-hygroscopic” material has a water content of less thanabout 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%,0.2%, or 0.1% water by weight.

Advantageously, the free acid can be used to make both a solidformulation and an intravenous (IV) formulation. During the evaluation,it was found that the disodium salt, although unsuitable for solidcompositions such as tablets, was very soluble and therefore suitablefor IV formulations. Thus, in another embodiment, a sterile lyophilizedpowder for injection is made by forming a disodium salt in situ withsodium hydroxide and lyophilizing the resulting solution. The disodiumsalt is highly soluble and therefore is advantageous to reconstitute insterile water to yield a solution. In some embodiments, the resultingsolution may be added to an intravenous bag. The bag may contain anisotonic solution such as 0.9% sodium chloride or 5% dextrose.

In some embodiments, the salt solution, such as a disodium or monosodiumsalt, can be lyophilized by freezing the solution in a lyophilizer toabout −50 to −30° C. at about 0.1 to 1 degree/minute and holding it forabout 200-700 minutes at which point the chamber in the lyophilizer isevacuated to approximately 200-250 millitorr and the temperature isramped up to about −30 to about −10° C. at about 0.5 to about 3degrees/minute. The product is held at −30 to about −10° C. for about1000-2500 minutes and then the temperature is ramped up to about 21-35°C. at about 0.1 to 1 degrees/minute and held for 1000-2500 minutes togive the finished product.

In embodiments of some preparation methods, the crystalline free acid(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂) can be prepared byacidification of an aqueous solution of the corresponding salt, such asthe disodium salt 1 (R═PO(ONa)₂).

Any salt of the free acid 1 (R═PO(OH)₂) can be used to regenerate thefree acid by acidification. In some embodiments, the salt is an alkalimetal or an alkaline earth metal. In other embodiments, the salt is analkali metal salt, such as a disodium salt of 1 (R═PO(OH)₂).

It was found that the choice of acid is not critical. Any acid that issufficiently acidic to doubly protonate the phosphate disodium salt 1(R═PO(ONa)₂), or other salt, to yield the free acid 1 (R═PO(OH)₂) can beused. In some embodiments, the acid is HCl, HBr, or H₂SO₄.

After dissolving the salt of the (R)-3-(4-(2-(2-methyl

tetrazol-5-yl)-pyridin-5-yl)-3-fluoro-phenyl)-5-hydroxy-methyloxazolidin-2-one dihydrogen phosphate, and after, acidifying the saltsolution to form crystals, the crystals may be filtered from thesupernatant. In some embodiments, wet crystals may be dried, for exampleby using a vacuum or lyophilizing the crystals.

In some embodiments, crystalline refers to uniformly crystallinematerial of crystalline(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate, such as substantially purecrystals.

The terms “approximately, “about,” and “substantially” as used hereinrepresent an amount close to the stated amount that still performs thedesired function or achieves the desired result. For example, the terms“approximately,” “about” and “substantially” may refer to an amount thatis within less than 10% of, within less than 5% of, within less than 1%of, within less than 0.1% of, and within less than 0.01% of the statedamount.

For instance, in the pharmaceutical industry, it standard practice toprovide substantially pure material when formulating pharmaceuticalcompositions. Therefore, in some embodiments, “substantially pure”refers to the amount of purity required for formulating pharmaceuticals,which may include, for example, a small amount of amorphous material orother material, wherein the material may still achieve sufficientpourability, lack of hygroscopicity, and purity suitable forpharmaceutical use. In some embodiments, the crystalline free acid thatis substantially pure contains at least about 96% crystalline free acidby weight, such as at least about 96.1%, 96.2%, 96.3%, 96.4%, 96.5%,96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%,97.6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%,98.6%, 98.7%, 98.8%, 98.9%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%,99.6%, 99.7%, 99.8%, 99.9%, or 100% crystalline free acid by weight. Insome embodiments, the di- or mono-sodium salt in formulations describedherein have at least about 96%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%,96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%,97.6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%,98.6%, 98.7%, 98.8%, 98.9%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%,99.6%, 99.7%, 99.8%, 99.9%, or 100% crystalline salt by weight. Informulating pharmaceuticals, it is useful to provide a non-sticky solidcrystalline(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate that can be poured and accuratelyweighed for use in, for example, tablets and capsules. Therefore, insome embodiments, the crystalline material is in a pourable form suchthat the particles do not strongly adhere to each other or the vessel inwhich it is contained, such that it is capable of uniformly and steadilyflowing from a vessel.

Preparation of the free acid(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂), and of its disodiumsalt 1 (R═PO(ONa)₂) is described in US Patent Publ. No. 2007/0155798 andU.S. patent application Ser. No. 12/577,089, the latter of which isassigned to the same assignee as in the present application.

In embodiments of some preparation methods, the crystalline free acid(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂) can be prepared byacidification of an aqueous solution of the corresponding salt, such asthe disodium salt 1 (R═PO(ONa)₂).

Any salt of the free acid 1 (R═PO(OH)₂) can be used to regenerate thefree acid by acidification. In some embodiments, the salt is an alkalimetal or an alkaline earth metal. In other embodiments, the salt is analkali metal salt, such as a disodium salt of 1 (R═PO(OH)₂).

In additional embodiments of some preparation methods, the free aciditself can be used to prepare the crystalline form by dissolution in adissolution solvent, such as a dipolar aprotic solvent, for exampledimethyl sulfoxide (DMSO) or 1-methyl-2-pyrrolidone (NMP) followed byaddition of a crystallization-inducing solvent such as ethanol, acetone,acetonitrile, dioxane, heptanes, isopropyl alcohol, methanol,tetrahydrofuran, toluene, water, dichloromethane, methyl isobutyl ketoneand ethyl acetate. In some embodiments, the dissolution and thecrystallization-inducing solvents can be either a pure solvent or amixture of pure solvents, and can be either in the form of a liquid, avapor, or a second layer. In some embodiments of the latter two cases,the crystallization-inducing solvent can be employed according to thevapor diffusion method of growing crystals, or the solvent-layeringmethod, both of which are well-known to those of skill in the art.

In further embodiments of some preparation methods, the free acid can bedissolved in at least one dipolar aprotic solvent such as DMSO or NMP atan elevated temperature, and crystalline free acid 1 (R═PO(OH)₂)obtained by cooling of the resulting solution, according to methodswell-known to those of skill in the art. The solvent can either be pure,or itself a mixture of pure solvents

In formulating pharmaceuticals, it is useful to provide a solidcrystalline compound that can be easily formed into dosage forms, forexample, tablets. In addition, it is useful to shorten the length oftime necessary to make a compound. To address these needs, in someembodiments, a method of making crystalline 1 (R═PO(OH)₂) that resultsin increased particle size are disclosed that significantly decrease thefiltering time caused by fine particles that slow down the filteringstep. In further embodiments, crystalline 1 (R═PO(OH)₂) has a particularparticle size distribution, for example, that directly results from themethod without relying on sieving the material solely to obtain theparticle size distribution.

To this end, in some embodiments, the resulting larger particle size ofthe crystalline 1 (R═PO(OH)₂) may be made by a high temperatureprecipitation procedure. In addition, in embodiments wherein an acid isused to form the free acid from the salt, it was found that theincreasing the rate at which the reaction mixture was added to the acidaffects the particle size and makes the particles larger. Thus, in someembodiments, the reaction mixture may be contacted to the acid solutionas fast as possible, such that there is essentially immediate contactwith the acid solution. In conventional methods, the reaction mixturemade contact with the acid solution more slowly, because the acidsolution was added to the reaction mixture and therefore the reactionmixture may not contact the acid solution until some time after additionof the acid solution, causing much smaller particle size. It was foundthat reversing the step, that is, adding the reaction mixture to theacid solution, will allow the reaction mixture to effectivelyimmediately contact the acid over the course of introducing the reactionmixture to the acidic solution, which results in larger particle sizematerial. Thus, in some embodiments, immediate contact is made by addingthe reaction mixture to the acid solution. The reaction mixture may bepumped into the acid solution over time, for example, over a few hours,such as 1-4 hours.

In some embodiments, an aqueous ethanol- or THF-containing solution ofTR-701FA may be prepared by adding a sodium bicarbonate solution, forexample, a 2-10% solution by weight, such as a 5% solution. In someembodiments, the solution may be added to an aqueous acidic solution andethanol or THF to form the free acid. In some embodiments, from about0.5-10, about 1.5-3.0, or about 2.2 equivalents of 1M HCl may be used.In addition, in some embodiments, about 1-10 volumes, about 2-6 volumes,or about 4 volumes of ethanol may be used. THF may also be used. In someembodiments, the solution including the hydrochloric acid and ethanolmay be maintained at about 40-100° C., about 60-70° C., or about 65 to70° C. The acid and alcohol may be adjusted. The TR-701FA crystallizedduring this addition with a reduced amount of fines in the product incomparison to previously disclosed methods.

In some embodiments, the ethanol or THF prevents the free acid fromgelling during the process.

Typical particle size distribution is measured using a laser diffractionparticle size analyzer, namely a Malvern Mastersizer. D10 (μm)represents the diameter below which lies 10% of the total particlevolume. D50 (μm) is the median volume diameter. D90 (μm) is the diameterbelow which lies 90% of the total particle volume.

In some embodiments, when the particle size is not controlled, 10% ofthe total particle volume may have a diameter of less than about 0.28μm, the median volume diameter may be about 0.79 μm, and 90% of thetotal particle volume may have a diameter of less than about 0.44 μm. Bycontrolling (increasing) the particle size using methods disclosedherein, the particles are significantly larger overall.

In some embodiments, when the particle size is controlled using themethods described herein to increase particle size, 10% of the totalparticle volume may have an average diameter of at least about 0.5 μm,and/or the median volume diameter may be at least about 1.0 μm, and/or90% of the total particle volume may have an average diameter of atleast about 45 μm. In some embodiments, when the particle size iscontrolled (to increase particle size), 10% of the total particle volumemay have an average diameter of about 0.5-10 μm, such as about 1-5 μm.For example, when the particle size is controlled (to increase particlesize), 10% of the total particle volume may have an average diameter ofabout 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7,1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1,3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5,4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9,6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3,7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7,8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or 10.0 μm.

In some embodiments, when the particle size is controlled (to increaseparticle size), the median volume diameter may be greater than about 1.0μm, and have an average median volume diameter about 1-44 μm, about 1-40μm, about 10-35 μm, about 20-30 μm, or about 25-29, such as about 27 μm.In some embodiments, when the particle size is controlled to increaseparticle size, the average median volume diameter may be about 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 38, 39, 40, 41,42, 43, or 44 μm. For example, the average median volume diameter may beabout 25, 25.1, 25.2, 25.3, 25.4, 25.5, 25.6, 25.7, 25.8, 25.9, 26,26.1, 26.2, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27, 27.1, 27.2,27.3, 27.4, 27.5, 27.6, 27.7, 27.8, 27.9, 28, 28.1, 28.2, 28.3, 28.4,28.5, 28.6, 28.7, 28.8, 28.9, or 29 μm.

In some embodiments, when the particle size is controlled (to increaseparticle size), 90% of the total particle volume may have an averagediameter of the least about 45 μm such as about 45-100, about 45-80,about 55-75, or about 64-68 such as about 66. In some embodiments, whenthe particle size is controlled, 90% of the total particle volume mayhave an average diameter of about 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 μm. For example, 90% ofthe total particle volume may have an average diameter of about 64,64.1, 64.2, 64.3, 64.4, 64.5, 64.6, 64.7, 64.8, 64.9, 65, 65.1, 65.2,65.3, 65.4, 65.5, 65.6, 65.7, 65.8, 65.9, 66, 66.1, 66.2, 66.3, 66.4,66.5, 66.6, 66.7, 66.8, 66.9, 67, 67.1, 67.2, 67.3, 67.4, 67.5, 67.6,67.7, 67.8, 67.9, or 68 μm.

The crystalline free acid 1 (R═PO(OH)₂) may be characterized in havingthe FT-Raman for example as shown in FIG. 1, and the X-ray powderdiffraction for example as shown in FIG. 2, with the correspondingnumerical data for example as shown in Table 1 and Table 2 respectively.FIG. 3, FIG. 4, FIG. 5 and FIG. 6 show examples of the differentialscanning calorimetry (DSC) thermogram, solution ¹H NMR spectrum, theTG-FTIR diagram, and the dynamic vapor sorption (DVS) behavior ofcrystalline 1 (R═PO(OH)₂) respectively.

TABLE 1 FT-Raman spectroscopic data for crystalline free acid 1 (R =PO(OH)₂) Wavenumber (cm-1) Absolute Intensity 1612.9 2.57 1579.0 0.381521.3 0.25 1455.8 0.26 1404.9 0.39 1324.4 0.82 1274.7 0.24 1149.9 0.171018.3 0.22

TABLE 2 X-ray powder pattern diffraction data for crystalline free acid1 (R = PO(OH)₂) Angle 2Theta/° Intensity/% 10.6 17 13.7 6 13.9 8 14.7 3815.2 28 15.4 28 15.7 16 16.6 65 17.1 10 19.2 19 20.3 100 21.4 25 22.4 2323.2 8 23.6 7 24.7 29 25.3 14 25.9 8 26.8 82 28.2 44 28.4 24 29.0 8 30.38 30.8 11 31.0 13 31.9 8 33.5 17 34.7 7

In some embodiments, the distinguishing peaks for the crystalline freeacid comprise the following peaks: 14.7°, 15.2°, 16.6°, 20.3°, 26.8°,and 28.2°.

In other embodiments, the distinguishing peaks for the crystalline freeacid comprise the following peaks: 10.6°, 13.9°, 14.7°, 15.2°, 16.6°,20.3°, 26.8°, and 28.2°.

In some embodiments, the crystalline free acid comprises impurities thatare present in less than 1% of the purified crystalline free acid. Theseimpurities include

i.e.,5R)-3-{3-Fluoro-4-[6-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)-pyridin-3-yl]-phenyl}-5-hydroxymethyl-1,3-oxazolidin-2-one(“TR-700”) and/or

i.e.,(R)-5-(chloromethyl)-3-(3-fluoro-4-(6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl)phenyl)oxazolidin-2-one(“chloro impurity”).

Of the conventionally produced material having impurities that wereidentified using HPLC in Example 15, at least 2% by weight of the chloroimpurity was present. In purified crystalline free acid made using themethod of making the free acid disclosed in U.S. patent application Ser.No. 12/577,089, which is assigned to the same assignee as in the presentapplication, and the crystallization methods disclosed herein, thechloro impurity was present in less than about 1% by weight, such asless than about 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1%by weight of the of the crystalline free acid. In some embodiments thechloro impurity may be reduced to much lower than 0.1% by weight, suchas, less than about 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%,0.02%, or 0.01% by weight of the crystalline free acid. In someembodiments, the purified crystalline free acid is substantially free ofthe chloro impurity.

Of the conventionally produced material having impurities that wereidentified using HPLC in Example 15, at least about 1% by weight of theTR-700 impurity was present. In purified crystalline free acid madeusing the method of making the free acid disclosed in U.S. patentapplication Ser. No. 12/577,089, which is assigned to the same assigneeas in the present application, and the crystallization methods disclosedherein, the TR-700 impurity was present in less than about 1% by weight.In some embodiments, the crystalline free acid contains less than about0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% by weight of theTR-700 impurity. In some embodiments, the crystalline free acid issubstantially free of the TR-700 impurity.

In addition, purified crystalline free acid made using the method ofmaking the free acid disclosed in U.S. patent application Ser. No.12/577,089, which is assigned to the same assignee as in the presentapplication, and the crystallization methods disclosed herein, may alsobe distinguished from the conventionally produced crystalline free acidby the presence of the following compounds. For example, the followingimpurities were not found in a sample of conventionally producedcrystalline free acid as shown in Example 15:

(hereinafter “des-methyl TR-701”), i.e., dihydrogen((5R)-3-{3-fluoro-4-[6-(2H-1,2,3,4-tetrazol-5-yl)-3-pyridinyl]phenyl}-2-oxo-1,3-oxazolan-5-yl)methylphosphate;

(hereinafter “overalkylated-phosphorylated impurity”), i.e.,51-43-(3-fluoro-4-(6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl)phenyl)-2-oxooxazolidin-5-yl)methoxy)-3-hydroxypropan-2-yldihydrogen phosphate and,3-((3-(3-fluoro-4-(6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl)phenyl)-2-oxooxazolidin-5-yl)methoxy)-2-hydroxypropyldihydrogen phosphate;

(hereinafter “one of the OA-700 mixed di ester”) i.e.,3-{[(5R)-3-{3-fluoro-4-[6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl]phenyl}-2-oxo-1,3-oxazolidin-5-yl]methoxy}-2-hydroxypropyl[(5R)-3-{3-fluoro-4-[6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl]phenyl}-2-oxo-1,3-oxazolidin-5-yl]methylhydrogen phosphate; and/or

(hereinafter “another of the OA-700 mixed di ester”) i.e.,2-{[(5R)-3-{3-fluoro-4-[6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl]phenyl}-2-oxo-1,3-oxazolidin-5-yl]methoxy}-1-hydroxyethyl[(5R)-3-{3-fluoro-4-[6-(2-methyl-2H-tetrazol-5-yl)pyridin-3-yl]phenyl}-2-oxo-1,3-oxazolidin-5-yl]methylhydrogen phosphate.

Those skilled in the art will appreciate that variousisotopically-substituted variants (through, e.g., substitution ofdeuterium for hydrogen, ¹³C for carbon, ¹⁵N for nitrogen, or ³²P forphosphorus) can also be readily produced. All such variants arecontemplated within the scope of this disclosure.

In various embodiments, the purified crystallized free acid disclosedherein can be used alone, in combination with other compounds disclosedherein, or in combination with one or more other agents active in thetherapeutic areas described herein.

In another aspect, the present disclosure relates to a pharmaceuticalcomposition comprising one or more physiologically acceptable surfaceactive agents, additional carriers, diluents, excipients, smoothingagents, suspension agents, film forming substances, and coatingassistants, or a combination thereof; and a composition disclosedherein. Acceptable additional carriers or diluents for therapeutic useare well known in the pharmaceutical art, and are described, forexample, in Remington's Pharmaceutical Sciences, 18th Ed., MackPublishing Co., Easton, Pa. (1990), which is incorporated herein byreference in its entirety. Preservatives, stabilizers, dyes, sweeteners,fragrances, flavoring agents, and the like may be provided in thepharmaceutical composition. For example, sodium benzoate, ascorbic acidand esters of p-hydroxybenzoic acid may be added as preservatives. Inaddition, antioxidants and suspending agents may be used. In variousembodiments, alcohols, esters, sulfated aliphatic alcohols, and the likemay be used as surface active agents; sucrose, glucose, lactose, starch,microcrystalline cellulose, crystallized cellulose, mannitol, lightanhydrous silicate, magnesium aluminate, magnesium metasilicatealuminate, synthetic aluminum silicate, calcium carbonate, sodium acidcarbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose,and the like may be used as excipients; magnesium stearate, talc,hardened oil and the like may be used as smoothing agents; coconut oil,olive oil, sesame oil, peanut oil, soya may be used as suspension agentsor lubricants; cellulose acetate phthalate as a derivative of acarbohydrate such as cellulose or sugar, or methylacetate-methacrylatecopolymer as a derivative of polyvinyl may be used as suspension agents;and plasticizers such as ester phthalates and the like may be used assuspension agents.

The term “pharmaceutical composition” refers to a mixture of a compounddisclosed herein with other chemical components, such as diluents oradditional carriers. The pharmaceutical composition facilitatesadministration of the compound to an organism. Multiple techniques ofadministering a pharmaceutical composition exist in the art including,but not limited to, oral, injection, aerosol, parenteral, and topicaladministration. Pharmaceutical compositions can also be obtained byreacting the free acid with inorganic or organic bases such as sodiumhydroxide or magnesium hydroxide. In some embodiments, pharmaceuticallyacceptable salts of the compounds disclosed herein (e.g., as made insitu during the manufacture of an intravenous formulation) are provided.In some embodiments, sodium hydroxide is used to prepare a lyophilizedpowder the formulation that comprises a salt of the free acid, which isproduced in situ.

The term “carrier” refers to a chemical compound that facilitates theincorporation of a compound into cells or tissues.

The term “diluent” refers to chemical compounds diluted in water thatwill dissolve the composition of interest as well as stabilize thebiologically active form of the compound. Salts dissolved in bufferedsolutions are utilized as diluents in the art. One commonly usedbuffered solution is phosphate buffered saline because it mimics thesalt conditions of human blood. Since buffer salts can control the pH ofa solution at low concentrations, a buffered diluent rarely modifies thebiological activity of a compound. As used herein, an “excipient” refersto an inert substance that is added to a composition to provide, withoutlimitation, bulk, consistency, stability, binding ability, lubrication,disintegrating ability, etc., to the composition. A “diluent” is a typeof excipient.

The term “physiologically acceptable” refers to a carrier or diluentthat does not abrogate the biological activity and properties of thecompound.

The pharmaceutical compounds described herein can be administered to ahuman patient per se, or in pharmaceutical compositions where they aremixed with other active ingredients, as in combination therapy, orsuitable carriers or excipient(s). Techniques for formulation andadministration of the compounds of the instant application may be foundin “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton,Pa., 18th edition, 1990.

Suitable routes of administration may, for example, include oral,rectal, transmucosal, topical, or intestinal administration; parenteraldelivery, including intramuscular, subcutaneous, intravenous,intramedullary injections, as well as intrathecal, directintraventricular, intraperitoneal, intranasal, or intraocularinjections. The compound can also be administered in sustained orcontrolled release dosage forms, including depot injections, osmoticpumps, pills, transdermal (including electrotransport) patches, and thelike, for prolonged and/or timed, pulsed administration at apredetermined rate.

The pharmaceutical compositions of the present invention may bemanufactured in a manner that is itself known, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or tabletting processes.

Pharmaceutical compositions may be formulated in any conventional mannerusing one or more physiologically acceptable carriers comprisingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Properformulation is dependent upon the route of administration chosen. Any ofthe well-known techniques, diluents, carriers, and excipients may beused as suitable and as understood in the art; e.g., in Remington'sPharmaceutical Sciences, above.

Injectables can be prepared in conventional forms, either as liquidsolutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. Suitableexcipients are, for example, water, saline, dextrose, mannitol, lactose,lecithin, albumin, sodium glutamate, cysteine hydrochloride, and thelike. In addition, if desired, the injectable pharmaceuticalcompositions may contain minor amounts of nontoxic auxiliary substances,such as wetting agents, pH buffering agents, and the like.Physiologically compatible buffers include, but are not limited to,Hanks's solution, Ringer's solution, or physiological saline buffer. Ifdesired, absorption enhancing preparations may be utilized.

For transmucosal administration, penetrants appropriate to the barrierto be permeated may be used in the formulation.

Pharmaceutical formulations for parenteral administration, e.g., bybolus injection or continuous infusion, include aqueous solutions of theactive compounds in water-soluble form. Additionally, suspensions of theactive compounds may be prepared as appropriate oily injectionsuspensions. Aqueous injection suspensions may contain substances whichincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents that increase the solubility ofthe compounds to allow for the preparation of highly concentratedsolutions. Formulations for injection may be presented in unit dosageform, e.g., in ampoules or in multi-dose containers, with an addedpreservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

For oral administration, the composition can be formulated readily bycombining the compositions of interest with pharmaceutically acceptablecarriers well known in the art. Such carriers, which may be used inaddition to the cationic polymeric carrier, enable the compositions ofthe invention to be formulated as tablets, pills, dragees, capsules,liquids, gels, syrups, slurries, suspensions and the like, for oralingestion by a patient to be treated. Pharmaceutical preparations fororal use can be obtained by combining the active compounds with solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose, and/or polyvinylpyrrolidone (PVP), e.g.,Povidone. If desired, disintegrating agents may be added, such as thecross-linked polyvinylpyrrolidone (e.g. Crospovidone), agar, or alginicacid or a salt thereof such as sodium alginate. Dragee cores areprovided with suitable coatings. For this purpose, concentrated sugarsolutions may be used, which may optionally contain gum arabic, talc,polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/ortitanium dioxide, lacquer solutions, and suitable organic solvents orsolvent mixtures. Dyestuffs or pigments may be added to the tablets ordragee coatings for identification or to characterize differentcombinations of active compound doses. For this purpose, concentratedsugar solutions may be used, which may optionally contain gum arabic,talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/ortitanium dioxide, lacquer solutions, and suitable organic solvents orsolvent mixtures. Dyestuffs or pigments may be added to the tablets ordragee coatings for identification or to characterize differentcombinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. All formulations fororal administration should be in dosages suitable for suchadministration.

For buccal administration, the compositions may take the form of tabletsor lozenges formulated in conventional manner.

For administration by inhalation, the composition can be convenientlydelivered in the form of an aerosol spray presentation from pressurizedpacks or a nebulizer, with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol the dosage unit may be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof, e.g., gelatin for use in an inhaler or insufflator may be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

Further disclosed herein are various pharmaceutical compositions wellknown in the pharmaceutical art for uses that include intraocular,intranasal, and intraauricular delivery. Suitable penetrants for theseuses are generally known in the art. Such suitable pharmaceuticalformulations are most often and preferably formulated to be sterile,isotonic and buffered for stability and comfort. Pharmaceuticalcompositions for intranasal delivery may also include drops and spraysoften prepared to simulate in many respects nasal secretions to ensuremaintenance of normal ciliary action. As disclosed in Remington'sPharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, Pa.(1990), which is incorporated herein by reference in its entirety, andwell-known to those skilled in the art, suitable formulations are mostoften and preferably isotonic, slightly buffered to maintain a pH of 5.5to 6.5, and most often and preferably include antimicrobialpreservatives and appropriate drug stabilizers. Pharmaceuticalformulations for intraauricular delivery include suspensions andointments for topical application in the ear. Common solvents for suchaural formulations include glycerin and water.

The compositions may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, the compositionsmay also be formulated as a depot preparation. Such long actingformulations may be administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection. Thus,for example, the compounds may be formulated with suitable polymeric orhydrophobic materials (for example as an emulsion in an acceptable oil)or ion exchange resins, or as sparingly soluble derivatives, forexample, as a sparingly soluble salt.

For hydrophobic compounds, a suitable pharmaceutical carrier may be acosolvent system comprising benzyl alcohol, a nonpolar surfactant, awater-miscible organic polymer, and an aqueous phase. A common cosolventsystem used is the VPD co-solvent system, which is a solution of 3% w/vbenzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™, and65% w/v polyethylene glycol 300, made up to volume in absolute ethanol.Naturally, the proportions of a co-solvent system may be variedconsiderably without destroying its solubility and toxicitycharacteristics. Furthermore, the identity of the co-solvent componentsmay be varied: for example, other low-toxicity nonpolar surfactants maybe used instead of POLYSORBATE 80™; the fraction size of polyethyleneglycol may be varied; other biocompatible polymers may replacepolyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars orpolysaccharides may substitute for dextrose.

Methods for treating bacterial infections may include administering atherapeutically effective amount of the therapeutic compounds asdescribed herein. Treating a bacterial infection may also includeprophylactically administering the therapeutic compounds to preventinfection or the spread of an infection in a subject at imminent risk ofinfection, such as a subject receiving or about to undergo surgery, animmunocompromised subject, or subject otherwise at risk of an infectionif the compound was not administered. The compounds show inhibitoryactivity against a broad spectrum of bacteria, against methicillinresistant Staphylococcus aureus (MRSA) and vancomycin resistantEnterococci (VRE) and have excellent relative antibiotic activity with arelatively low concentration thereof or in vivo. Further, the compoundsof the present invention may exert potent antibacterial activity versusvarious human and animal pathogens, including Gram-positive bacteriasuch as Staphylococi, Enterococci and Streptococi, anaerobicmicroorganisms such as Bacteroides and Clostridia, and acid-resistantmicroorganisms such as Mycobacterium tuberculosis and Mycobacteriumavium. In an embodiment, the bacterial infection that may be treated orameliorated is MRSA.

The compositions or pharmaceutical compositions described herein may beadministered to the subject by any suitable means. Non-limiting examplesof methods of administration include, among others, (a) administrationthough oral pathways, which administration includes administration incapsule, tablet, granule, spray, syrup, or other such forms; (b)administration through non-oral pathways such as rectal, vaginal,intraurethral, intraocular, intranasal, or intraauricular, whichadministration includes administration as an aqueous suspension, an oilypreparation or the like or as a drip, spray, suppository, salve,ointment or the like; (c) administration via injection, subcutaneously,intraperitoneally, intravenously, intramuscularly, intradermally,intraorbitally, intracapsularly, intraspinally, intrasternally, or thelike, including infusion pump delivery; as well as (d) administrationtopically; as deemed appropriate by those of skill in the art forbringing the active compound into contact with living tissue.

Pharmaceutical compositions suitable for administration includecompositions where the active ingredients are contained in an amounteffective to achieve its intended purpose. The therapeutically effectiveamount of the compounds disclosed herein required as a dose will dependon the route of administration, the type of animal, including human,being treated, and the physical characteristics of the specific animalunder consideration. The dose can be tailored to achieve a desiredeffect, but will depend on such factors as weight, diet, concurrentmedication and other factors which those skilled in the medical artswill recognize. More specifically, a therapeutically effective amountmeans an amount of compound effective to prevent, alleviate orameliorate symptoms of disease or prolong the survival of the subjectbeing treated. Determination of a therapeutically effective amount iswell within the capability of those skilled in the art, especially inlight of the detailed disclosure provided herein.

As will be readily apparent to one skilled in the art, the useful invivo dosage to be administered and the particular mode of administrationwill vary depending upon the age, weight and mammalian species treated,the particular compounds employed, and the specific use for which thesecompounds are employed. The determination of effective dosage levels,that is the dosage levels necessary to achieve the desired result, canbe accomplished by one skilled in the art using routine pharmacologicalmethods. Typically, human clinical applications of products arecommenced at lower dosage levels, with dosage level being increaseduntil the desired effect is achieved. Alternatively, acceptable in vitrostudies can be used to establish useful doses and routes ofadministration of the compositions identified by the present methodsusing established pharmacological methods.

In non-human animal studies, applications of potential products arecommenced at higher dosage levels, with dosage being decreased until thedesired effect is no longer achieved adverse side effects disappear. Thedosage may range broadly, depending upon the desired effects and thetherapeutic indication. Typically, dosages may be about 10 microgram/kgto about 100 mg/kg body weight, preferably about 100 microgram/kg toabout 10 mg/kg body weight. Alternatively dosages may be based andcalculated upon the surface area of the patient, as understood by thoseof skill in the art.

The exact formulation, route of administration and dosage for thepharmaceutical compositions of the present invention can be chosen bythe individual physician in view of the patient's condition. (See e.g.,Fingl et al. 1975, in “The Pharmacological Basis of Therapeutics”, whichis hereby incorporated herein by reference in its entirety, withparticular reference to Ch. 1, p. 1). Typically, the dose range of thecomposition administered to the patient can be from about 0.5 to about1000 mg/kg of the patient's body weight. The dosage may be a single oneor a series of two or more given in the course of one or more days, asis needed by the patient. In instances where human dosages for compoundshave been established for at least some condition, the present inventionwill use those same dosages, or dosages that are about 0.1% to about500%, more preferably about 25% to about 250% of the established humandosage. Where no human dosage is established, as will be the case fornewly-discovered pharmaceutical compositions, a suitable human dosagecan be inferred from ED₅₀ or ID₅₀ values, or other appropriate valuesderived from in vitro or in vivo studies, as qualified by toxicitystudies and efficacy studies in animals.

It should be noted that the attending physician would know how to andwhen to terminate, interrupt, or adjust administration due to toxicityor organ dysfunctions. Conversely, the attending physician would alsoknow to adjust treatment to higher levels if the clinical response werenot adequate (precluding toxicity). The magnitude of an administrateddose in the management of the disorder of interest will vary with theseverity of the condition to be treated and to the route ofadministration. The severity of the condition may, for example, beevaluated, in part, by standard prognostic evaluation methods. Further,the dose and perhaps dose frequency will also vary according to the age,body weight, and response of the individual patient. A programcomparable to that discussed above may be used in veterinary medicine.

Although the exact dosage will be determined on a drug-by-drug basis, inmost cases, some generalizations regarding the dosage can be made. Thedaily dosage regimen for an adult human patient may be, for example, anoral dose of about 0.1 mg to 2000 mg of each active ingredient,preferably about 1 mg to about 500 mg, e.g. 5 to 200 mg. In otherembodiments, an intravenous, subcutaneous, or intramuscular dose of eachactive ingredient of about 0.01 mg to about 100 mg, preferably about 0.1mg to about 60 mg, e.g. about 1 to about 40 mg is used. In cases ofadministration of a pharmaceutically acceptable salt, dosages may becalculated as the free base. In some embodiments, the composition isadministered 1 to 4 times per day. Alternatively the compositions of theinvention may be administered by continuous intravenous infusion,preferably at a dose of each active ingredient up to about 1000 mg perday. As will be understood by those of skill in the art, in certainsituations it may be necessary to administer the compounds disclosedherein in amounts that exceed, or even far exceed, the above-stated,preferred dosage range in order to effectively and aggressively treatparticularly aggressive diseases or infections. In some embodiments, thecompounds will be administered for a period of continuous therapy, forexample for a week or more, or for months or years.

Dosage amount and interval may be adjusted individually to provideplasma levels of the active moiety which are sufficient to maintain themodulating effects, or minimal effective concentration (MEC). The MECwill vary for each compound but can be estimated from in vitro data.Dosages necessary to achieve the MEC will depend on individualcharacteristics and route of administration. However, HPLC assays orbioassays can be used to determine plasma concentrations.

Dosage intervals can also be determined using MEC value. Compositionsshould be administered using a regimen which maintains plasma levelsabove the MEC for 10-90% of the time, preferably between 30-90% and mostpreferably between 50-90%.

In cases of local administration or selective uptake, the effectivelocal concentration of the drug may not be related to plasmaconcentration.

The amount of composition administered may be dependent on the subjectbeing treated, on the subject's weight, the severity of the infection,the manner of administration and the judgment of the prescribingphysician.

Compositions disclosed herein can be evaluated for efficacy and toxicityusing known methods. For example, the toxicology of the compound may beestablished by determining in vitro toxicity towards a cell line, suchas a mammalian, and preferably human, cell line. The results of suchstudies are often predictive of toxicity in animals, such as mammals, ormore specifically, humans. Alternatively, the toxicity of particularcompounds in an animal model, such as mice, rats, rabbits, or monkeys,may be determined using known methods. The efficacy of a particularcompound may be established using several recognized methods, such as invitro methods, animal models, or human clinical trials. Recognized invitro models exist for nearly every class of condition. Similarly,acceptable animal models may be used to establish efficacy of chemicalsto treat such conditions. When selecting a model to determine efficacy,the skilled artisan can be guided by the state of the art to choose anappropriate model, dose, and route of administration, and regime. Ofcourse, human clinical trials can also be used to determine the efficacyof a compound in humans.

The compositions may, if desired, be presented in a pack or dispenserdevice which may contain one or more unit dosage forms containing theactive ingredient. The pack may for example comprise metal or plasticfoil, such as a blister pack. The pack or dispenser device may beaccompanied by instructions for administration. The pack or dispensermay also be accompanied with a notice associated with the container inform prescribed by a governmental agency regulating the manufacture,use, or sale of pharmaceuticals, which notice is reflective of approvalby the agency of the form of the drug for human or veterinaryadministration. Such notice, for example, may be the labeling approvedby the U.S. Food and Drug Administration for prescription drugs, or theapproved product insert. Compositions comprising a compound of theinvention formulated in a compatible pharmaceutical carrier may also beprepared, placed in an appropriate container, and labeled for treatmentof an indicated condition.

A. Examples 1. Instrumentation

Raman microscopy was performed on a Renishaw System 1000, withstabilized diode laser 385 nm excitation and a NIR enhancedPeltier-cooled charge coupled device camera as detector. Measurementswere carried out with 50× or a long working distance 20× objective overa frequency range of 2000-100 cm⁻¹.

FT-Raman spectra were obtained on a Bruker RFS100 spectrometer withNd:YAG 1064 nm excitation, 100 mW laser power, and a Ge detector.Sixty-four scans were recorded over the range 25-3500 cm⁻¹, at 2 cm⁻¹resolution.

Bruker D8; Bragg-Brentano, reflection geometry; Copper K(alpha)radiation, 40 kV/40 mA; variable divergence slit; LynxEye detector with3° window; step size, 0.02-°2; step time, 37 s. The samples were rotated(0.5 rps) during the measurement.

Sample preparation: The samples were generally prepared without anyspecial treatment other than the application of slight pressure to get aflat surface. Silicon single crystal sample holder types: a) standardholder for polymorph screening, 0.1 mm deep, less than 20 mg samplerequired; b) 0.5 mm deep, 12 mm cavity diameter, ca. 40 mg required; c)1.0 mm deep, 12 mm cavity diameter, ca. 80 mg required. Normally sampleswere measured uncovered. Kapton foil or PMMA “dome” covers are alwaysindicated on the diffractogram with the sample identification.

2. Preparationee Acid 1 (R═PO(OH)₂) Example 1

A solution of 1 (R═PO(ONa)₂) was prepared in H₂O and 1M HCl added togive a fine suspension, which, after addition of tetrahydrofuran (THF),was stirred and filtered. The resulting crystalline solid 1 (R═PO(OH)₂)was dried in vacuum, and characterized by FT-Raman (FTR) (FIG. 1), X-raypowder diffraction (XRPD, Malvern Mastersizer) (FIG. 2),thermogravimetry-Fourier transform infrared spectroscopy (TG-FTIR), anddifferential scanning calorimetry (DSC). DSC measurement showed amelting point at 256.9° C. followed by a decomposition of the sample(FIG. 3).

Example 2

To 1 (R═PO(ONa)₂) (2 g) dissolved in 10 mL H₂O was slowly added HCl (6mL; 1M) to yield a fine suspension of a light yellow solid. Afteraddition of a further 5 mL H₂O and 20 mL THF the suspension was filteredand dried in vacuum.

Example 3

To 1 (R═PO(ONa)₂) (2 g) dissolved in 10 mL H₂O was slowly added HCl (8mL; 1M) to give a fine suspension of a light yellow solid, to which afurther 25 mL H₂O were added. The solid was filtered, washed with 10 mL0.1M HCl and 100 mL water and dried in vacuum.

Example 4

To 1 (R═PO(ONa)₂) (5 g) dissolved in 30 mL water was added 15 mL HCl(1M) and 30 mL of THF to produce a light yellow suspension, which wasstirred 30 min at room temperature and filtered. The resulting solid wassuspended in 150 mL water and stirred 60 min at room temperature. Then50 mL THF were added and the suspension was stirred 18 h. The suspensionwas filtered and the solid was washed with 10 mL HCl (0.1M) and 100 mLwater and dried in vacuum (15 h).

Example 5

To 1 (R═PO(ONa)₂) (2 g) dissolved in 15 mL water was slowly added HCl (6mL; 1M) to give a light yellow suspension. After addition of 20 mL THFand 60 mL water the suspension was stirred 18 hours, filtered, and thesolid was stirred again in 6 mL HCl (1M) for 15 min. Afterwards thesuspension was filtered and the solid was dried in vacuum.

Example 6

To 1 (R═PO(ONa)₂) (3 g) dissolved in 35 mL water was added HCl (9 mL;1M) to yield a light yellow suspension. After addition of 20 mL THF thesuspension was stirred 30 min at room temperature and then filtered. Theresulting solid was washed with 20 mL HCl (0.1M) and water and dried invacuum.

Example 7

Solid dihydrogen phosphate is added to a volume of DMSO orN-methylpyrrolidinone at about 50° C. until no more salt dissolves. Thesolution containing suspended salt is then heated further just until theremaining solid dissolves, and the solution filtered while hot andallowed to cool undisturbed, when it deposits crystals of the dihydrogenphosphate.

Example 8

A solution of the dihydrogen phosphate is prepared in DMSO orN-methylpyrrolidinone and filtered. To the filtered solution is addedethanol with stirring until the solution becomes cloudy. Stirring isthen discontinued, and a layer of ethanol carefully placed on top of thecloudy solution, which is allowed to sit undisturbed, when it depositscrystals of the dihydrogen phosphate.

Example 9

A solution of the dihydrogen phosphate is prepared in DMSO orN-methylpyrrolidinone and filtered. The filtered solution is thenexposed to vapor of ethanol, for example by placing an open container ofthe solution and an open container of ethanol together in a sealedvessel such that the two containers share a common headspace inside thevessel. On standing the container with the solution deposits crystals ofthe dihydrogen phosphate.

Example 10

A solution of a salt of the dihydrogen phosphate, such as the mono- ordisodium phosphate, is prepared. Such a solution can be prepared by suchmethods as simply dissolving a sample of the solid disodium phosphate inwater, or by adding the dihydrogen phosphate to an aqueous solution of abase sufficiently strong to substantially deprotonate the dihydrogenphosphate. Identification of an appropriate base is a routine matter forthe practicing chemist. Typically the resulting solution of the salt ofthe dihydrogen phosphate is then filtered, and to the filtrate is addedan acid to reprotonate the salt and induce crystallization of thedihydrogen phosphate. In a typical example, the dihydrogen phosphate isadded to an aqueous solution containing NaOH or Na₂CO₃ to yield asolution of the disodium phosphate, to which after filtration is addedaqueous or gaseous HCl to regenerate the dihydrogen phosphate, whichdeposits as crystals.

For pharmaceutical purposes it is advantageous to usepharmaceutically-acceptable acids and bases in this process, such asthose compiled in Handbook of Pharmaceutical Salts Properties, Selectionand Use. (P. Heinrich Stahl and Camille G. Wermuth, eds.) InternationalUnion of Pure and Applied Chemistry, Wiley-VCH 2002 and L. D. Bighley,S. M. Berge, D. C. Monkhouse, in “Encyclopedia of PharmaceuticalTechnology’. Eds. J. Swarbrick and J. C. Boylan, Vol. 13, Marcel Dekker,Inc., New York, Basel, Hong Kong 1995, pp. 453-499 discusses such saltsin detail.

As those skilled in the art will appreciate, elements of the methodsabove can be combined. For example, a solution of the dihydrogenphosphate in DMSO or N-methylpyrrolidinone can be prepared at onetemperature, a second solvent such as ethanol added, and the resultingsolution allowed to cool. Similarly, mixtures of solvents can be usedinstead of pure solvents, as is well-known to those skilled incrystallizing compounds. Furthermore, other solvents and mixturesthereof can also be used.

Elemental analysis for C₁₇H₁₆FN₆O₆P (measured/calculated) C 43.9 (44.8);H 3.6 (3.7); N 18.1 (18.4); O 21.2 (22.1); F 4.2 (4.2); P 6.7 (6.8).

Example 11

The particle size was measured using a Malvern Mastersizer. The samplinginstructions that were consistent with the instrument manufacturer'sinstructions were followed. The sample was prepared by suspending in 1-2mL of deionized water and sonicating for 3 minutes.

An exemplary particle size distribution of crystalline material such asthose described in Examples 1-10 above is set forth in FIG. 10 and Table3 below:

TABLE 3 Typical Particle Size Distribution (uncontrolled process) Lot02090054 D10 (um) D50 (um) D90 (um) Average 0.28 0.79 44

Example 12 Particle-Size Adjustment Experimental

A 22-L reactor was charged with 1M HCl (1.95 L, 2.2 equivalents) andethanol (1.6 L, 4 volumes), and the solution was heated to 70° C. Aseparate 12-L reactor equipped with a gas bubbler to monitor gasevolution was charged with TR-701FA [0.4 kg, AMRI lot # DUG-AH-166(2)],water (2.8 L, 7 vol), and ethanol (0.4 L, 1 vol). The slurry was stirredat ambient temperature and 5 wt % aqueous NaHCO₃ was added viaperistaltic pump over 30 minutes. No foaming was observed, however thegas evolution was vigorous as observed through the gas bubbler. Uponcompletion of the addition, the clear yellow solution was pH 6.6. Theaqueous TR-701 solution was added via peristaltic pump to theethanol/HCl solution over 90 minutes. Upon completion of the addition,the pH of the reaction mixture was 1.9 and the reaction mixture wascooled to 30° C. A sample of the slurry was withdrawn for analysis byoptical microscopy. The slurry was filtered through a polypropylenefilter cloth and the reactor and filter cake were rinsed with water (5volumes) and acetone (5 volumes). The total filtration time includingthe washes was 12 minutes. The solids were dried under high vacuum at50° C. to afford 391.7 g of reprecipitated TR-701FA (98% yield).Analysis by ¹H NMR was consistent with the assigned structure. HPLCanalysis (Method A): 98.8% (AUC) t_(R)=5.2 min. The level of residualethanol by ¹H NMR analysis was 0.03%, the water content was 0.15% byKarl Fischer titration, and the sodium content was 5 ppm.

The particle size was measured using a Malvern Mastersizer laserscattering microscopy. The sampling instructions that were consistentwith the instrument manufacturer's instructions were followed. Thesample was prepared by suspending in 1-2 mL of deionized water andsonicating for 3 minutes. The laser diffraction data is set forth inFIG. 11 and in Table 4 below.

TABLE 4 Lot JAS-I-45 D10 (um) D50 (um) D90 (um) Average 0.45 14.13 38.42

In another experiment, the typical particle size distribution using acontrolled method, such as provided in this example, is set forth FIG.12 and in Table 5 below:

TABLE 5 Typical Particle Size Distribution (using particle size controlprocess) Lot 0209118 D10 (μm) D50 (μm) D90 (μm) Average 3.3 27 66 Rangefor Application 1-5 1-40 45-80

The immediate release formulation and the intravenous formulationdescribed in Examples 13-14 below were made using the crystalline freeacid wherein the particle size was controlled.

Example 13 Immediate Release Formulation

The qualitative and quantitative formulation of immediate release(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂) tablets (“TorezolidPhosphate Tablets”), 200 mg, is presented in Table 6. All componentsused in the manufacturing are listed with the quality standard,function, and weight percent of each individual component. The listingis inclusive of all materials used during the manufacture of the drugproduct whether or not they are present in the finished product.

TABLE 6 Composition of Torezolid Phosphate Tablets, 200 mg 200 mg TabletQuality Weight % Ingredient Standard Function (mg/unit) (w/w) TorezolidPhosphate¹ In-house Active Ingredient 200 50.0 MicrocrystallineCellulose NF Diluent 78.0 19.5 (Avicel PH-101) Mannitol² NF Diluent 78.019.5 (Mannogen ® EZ Spray Dried) Povidone NF Binder 16.0 4.0 (PlasdoneK-29/32) Crospovidone NF Disintegrant 24.0 6.0 (Kollidon ® CL) PurifiedWater² USP Granulating — — Medium Magnesium Stearate NF Lubricant 4.01.0 (HyQual ®) Vegetable Source Total Core Tablet Weight¹ 400.0 100.0Opadry II Yellow Colored Film 14.0 3.4 Coat Purified Water³ USP FilmCoating — — Medium Total Weight 414.0 103.4 Abbreviations: NF = NationalFormulary; USP = United States Pharmacopeia ¹The actual amount oftorezolid phosphate is adjusted based on potency of the drug substancelot used. ²The actual amount of mannitol is adjusted based on amount ofthe drug substance used. ³Removed during processing.

Example 14 Powder and Formulation for Injection

(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate 1 (R═PO(OH)₂) “) (“TorezolidPhosphate for Injection” or “TR-701 FA for Injection”), 200 mg/vial, wasprepared in a formulation as a sterile lyophilized powder for injection.TR-701 FA for Injection is formulated in situ as the disodium salt usingsodium hydroxide to take advantage of its superior aqueous solubility(>130 mg/mL).

TR-701 FA for Injection, 200 mg/vial, is to be reconstituted with 4 mLof Sterile Water for Injection (WFI), USP to yield a 50 mg/mL solution.The appropriate clinical dose volume is to be withdrawn from the vialand added to an intravenous (IV) non-di(2-ethylhexyl)phthalate (DEHP)bag containing either 0.9% Sodium Chloride Injection, USP (saline) or 5%Dextrose Injection, USP (dextrose). The resulting IV solution is to beinfused using a non-DEHP solution set with a 0.22 μm in-line filter.

The unit composition of TR-701 FA Compounding Solution forLyophilization is presented in Table 7 and the unit composition ofTR-701 FA for Injection, 200 mg/vial is presented in Table 8.

TABLE 7 Unit Composition of TR-701 FA Compounding Solution forLyophilization Component Function Theoretical Quantity TR-701 FA DrugSubstance 100 mg/mL Mannitol, Powder, USP Bulking Agent  50 mg/mL SodiumHydroxide, USP In-situ salt formation, qs for pH adjustment pHadjustment to 7.75 Hydrochloric Acid, NF pH adjustment qs for pHadjustment to 7.75 Water for Injection, USP/EP Manufacturing solvent qsto 1.0 mL

TABLE 8 Unit Composition of TR-701 FA for Injection, 200 mg/vialComponent Function Theoretical Quantity TR-701 FA Drug Substance 210 mg^(a) Mannitol, Powder, USP Bulking Agent 105 mg Sodium Hydroxide, USPIn-situ salt formation, qs for pH adjustment pH adjustment to 7.75Hydrochloric Acid, NF pH adjustment qs for pH adjustment to 7.75 Waterfor Injection, Manufacturing solvent qs to 2.1 mL USP/EP ^(b) ^(a) Avolume equivalent to 210 mg of TR-701 FA is filled into each vial to sothat reconstitution of the vial with 4.0 mL of Water for Injection (afinal volume of 4.2 mL is obtained due to volume displacement of thedissolved solids) results in a 50 mg/mL solution of TR-701 FA that willallow withdrawal of the label contents. ^(b) Water for Injection isessentially removed during lyophilization.

The typical manufacturing batch formula for TR-701 FA for Injection, 200mg/vial is presented in Table 9.

TABLE 9 Typical Batch Formula for TR-701 FA for Injection, 200 mg/vialMaterial Theoretical Quantity TR-701 FA 400 g ^(a) Mannitol, Powder, USP200 g Sodium Hydroxide, NF qs for pH adjustment to pH 7.75 HydrochloricAcid, NF qs for pH adjustment to pH 7.75 Water for Injection, USP/EP qs4276 g Total 4000 mL (~1900 vials) ^(a) The actual quantity of TR-701 FAdrug substance to be weighed is adjusted based on potency.

The manufacturing process is summarized below and schematics of theprocess for preparing a compounding solution and for sterile filtering,filling, and lyophilization are presented FIGS. 8 and 9.

Compounding Solution

The compounding solution is prepared in the following sequence:

Add approximately 50% of the total amount of Water for Injection to atared compounding vessel.

Add TR-701 FA and slowly neutralize with a solution of sodium hydroxidewhile mixing.

Add and dissolve mannitol with mixing.

Measure the pH of the resulting solution. If the solution is outside thetarget range of pH 7.70 to 7.80, adjust the pH using either 1N sodiumhydroxide or 1N hydrochloric acid.

Add Water for Injection to final volume and mix.

Sterile Filtering/Filling/Lyophilization

Filter the bulk solution through 2 integrity-tested 0.22 μm filters inseries and collect the solution in a sterile receiving vessel.

Add target fill weight of solution into 20 mL vials under asepticconditions.

Partially insert lyophilization stoppers into the vials.

Lyophilize the vials according to an appropriate cycle.

At the end of the lyophilization cycle, backfill the chamber withnitrogen and stopper vials under partial vacuum.

Seal vials with flip off caps.

Example 15

A sample of crystalline free acid which was made according to a methodof making the free acid disclosed in U.S. patent application Ser. No.12/577,089, which is assigned to the same assignee as in the presentapplication, and by using the crystallization methods described herein,was crystallized according to methods described herein was characterizedusing HPLC and contains various levels of impurities such as thosedescribed in Table 10 below:

TABLE 10 Identified Individual Impurities HPLC (TM.1911) NMT 0.5%Rx600013 NMT 0.5% Rx600024 NMT 0.5% Rx600014 NMT 0.2% Rx600023 NMT 0.5%Rx600025 NMT 0.5% Rx600020 NMT 2.0% Rx600001 NMT 1.5% Rx600022

In addition, a substantially pure sample of crystalline free acid whichwas made according to processes that were not disclosed in US PatentPublication No. 20070155798 and was crystallized according to methodsdescribed herein (hereinafter “ours”), was compared to a sample ofmaterial made by Dong-A Pharm. Co. (hereinafter “the Dong-A material”),which was given to Trius Therapeutics Inc. in approximately 2007. Thepotency of the Dong-A material was approximately 84% by weight of thesample in comparison to a substantially pure reference sample; however,the purity of the crystalline free acid was 94.1% by weight of thematerial identified by HPLC as indicated below. Therefore, approximately10% of the impurities in the Dong-A material was not identified by HPLC.The purity profile comparison is set forth in Table 11 below:

TABLE 11 Impurity Area Percent Name RRT Dong A ours ID 600011 0.54 0.12ND DA-1dimer diphos 600013** 0.56 ND 0.08 Des-Me UNK 0.65 0.07 ND UNK0.77 0.34 ND UNK 0.86-0.88 0.07 0.03 600024 0.91 0.22 0.12 PyrophosphateUNK 0.94 0.07 ND UNK 0.95 0.05 ND 600012 1 94.1 97.1 API 600023 1.080.14 ND N-1 Phosphorylated UNK 1.1 0.06 ND UNK 1.14 0.05 ND 600025**1.15 ND 0.27 Over-Alk'd pair UNK 1.2 0.07 ND UNK 1.21 0.05 ND UNK 1.310.05 ND UNK 1.39 0.26 0.04 UNK 1.47 0.35 ND 600020  1.5-1.51 0.2 0.08Dimer UNK 1.56 ~0.05 ND 600001*  1.67-1.688 1.12 0.63 TR-700 6000221.72-1.73 0.28 1.2 Bis 600042** 1.79 ND 0.12 OA-700 mixed diester600043** 1.8 ND 0.15 OA-700 mixed diester 600026* 2.27-2.28 2.17 0.06Chloro UNK 2.34 0.05 ND UNK 2.4 0.06 ND *equals ours << Dong A **equalsimpurity present in ours but not Dong A

Organic Impurities in TR-701 FA Drug Substance

Impurity ‘Name’ Structure and Chemical Name Rx600013 ‘Des-methyl TR-701’

Rx600024 ‘Pyrophosphate’

Rx600014 ‘Ring opened’

Rx600023 ‘Me-isomer’

Rx600025 ‘Overalkylated- phosphorylated impurity’

Rx600020 ‘Dimer impurity’

Rx600026 ‘Chloro’

Rx600001 TR-700

Rx600022 ‘Bis phosphate’

Rx600042

Rx600043

What is claimed is:
 1. Crystalline particles comprising at least about96% by weight of(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate wherein the remainder of thecrystalline particles comprises at least one compound selected from thegroup consisting of


2. The crystalline particles of claim 1, wherein the remainder of thecrystalline particles comprises at least one compound selected from thegroup consisting of:


3. The crystalline particles of claim 2 comprising at least about 97% byweight of(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen.
 4. The crystalline particles of claim 2,wherein the median volume diameter is at least about 1.0 μm.
 5. Apharmaceutical composition comprising the crystalline particles of claim1 and at least one pharmaceutically acceptable carrier, excipient ordiluent.
 6. A pharmaceutical composition comprising the crystallineparticles of claim 2 and at least one pharmaceutically acceptablecarrier, excipient or diluent.
 7. The pharmaceutical composition ofclaim 6, wherein the pharmaceutically acceptable carrier, excipient ordiluent is at least one member selected from the group consisting ofmannitol, polyvinylpyrrolidone, cross-linked polyvinylpyrrolidone, andmagnesium stearate.
 8. A reaction mixture comprising the crystallineparticles of claim 1 and a base.
 9. A reaction mixture comprising thecrystalline particles of claim 2 and a base.
 10. The reaction mixture ofclaim 8, wherein the base is sodium hydroxide.
 11. The reaction mixtureof claim 9, wherein the base is sodium hydroxide.
 12. A pharmaceuticalcomposition comprising a lyophilisate of the reaction mixture of claim8, comprising

wherein R═PO(ONa)₂.
 13. A pharmaceutical composition comprising alyophilisate of the reaction mixture of claim 9, comprising

wherein R═PO(ONa)₂.
 14. A pharmaceutical composition comprising acombination of at least about 96% by weight of a compound having thefollowing structure:

wherein R═PO(ONa)₂; and wherein the remainder of the combinationcomprises at least one salt of a compound selected from the groupconsisting of:

and at least one pharmaceutically acceptable carrier, excipient ordiluent.
 15. The pharmaceutical composition of claim 14, wherein thecombination comprises at least one salt of


16. The pharmaceutical composition of claim 14, wherein the combinationcomprises at least about 97% by weight of

wherein R═PO(ONa)₂.
 17. The pharmaceutical composition of claim 15wherein the combination comprises at least about 97% by weight of

wherein R═PO(ONa)₂.
 18. A method of treating a bacterial infectioncomprising administering an effective amount of the crystallineparticles of claim 1 to a subject in need thereof.
 19. A method oftreating a bacterial infection comprising administering an effectiveamount of the pharmaceutical composition of claim 15 to a subject inneed thereof.
 20. A process for making the crystalline particles ofclaim 1, comprising drying the crystalline particles.
 21. The process ofclaim 20, further comprising filtering the crystalline particles from asupernatant before the drying step.
 22. The process of claim 21, furthercomprising contacting a salt of crystalline(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate with an acid solution to formcrystallized(R)-3-(4-(2-(2-methyl-tetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one dihydrogen phosphate before the filtering step.
 23. Theprocess of claim 22, wherein the acid solution comprises HCl andethanol, or HCl and THF.